Ezup cfdna extraction kit

All cells were cultured in a humidified atmosphere of 5% CO₂ at 37°C. Two cell lines (MCF-7 and Hela) were cultured in a MEM media supplemented with 10% FBS, while cell line HEK293T was cultured in a high-glucose DMEM media. The cell-free DNA (cfDNA) of HEK293T, Hela and MCF-7 cells (5 × 10⁵/ml, 6 × 10⁵/ml and 7 × 10⁵/ml) were extracted using the Ezup cfDNA Extraction Kit (34 (link)) (Sangon Biotech Co. Ltd, China) according to the manufacturer's instruction. The extracted cfDNA was routinely stored at −80°C before incubation with our detection system.

LbCas12a (Cpf1), AsCas12a, FnCas12a and RNA inhibitor were purchased from New England Biolabs Co. Ltd (NEB, USA). N,N,N',N'-Tetramethylethylenediamine (TEMED), urea, ammonium persulphate (APS), 40% acrylamide/bis-acrylamide (19:1), Tris borate EDTA (TBE), 2 × TBE–urea sample buffer and Ezup cfDNA Extraction Kit were obtained from Sangon Biotech Co. Ltd (Shanghai, China). Dulbecco's modified Eagle's medium (DMEM), Minimum Essential Medium (MEM) and fetal bovine serum (FBS) were purchased from Invitrogen (Gibco, USA). Trypsin and dimethyl sulfoxide (DMSO) were purchased from Sigma-Aldrich (St Louis, MO, USA). Human breast cancer cell line (MCF-7), human cervical cancer cell line (HeLa) and human embryonic kidney cancer cell line (HEK293T) were obtained from the Cell Bank of Type Culture Collection of the Chinese Academy of Sciences (Shanghai, China). To create and maintain a ribonuclease-free environment, all the solutions containing RNAs were prepared with RNase-free water.